I would appreciate if anyone shared a thought on registering the
expressed Gal4- Binding Domain by Western blotting. I have used a
monoclonal Gal4-BD antibody by Clontech and at best I get unspecific
signals all over, with perhaps a stronger band near 22 kD in positive
control (but not entirely non-existant signal in the negative control,
only weaker than the positive). The isolated proteins are from Y190
strain transformed with Gal 4-BD , and PGBT9 plasmid was used. People
from Clontech claim that the promoter in this plasmid is too weak to get
a sufficient ammount of my protein.
Thanks,
Vuk
savv at medizin.uni-leipzig.de
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